Rapid, high-throughput DNA electrophoresis instruments are under intense scientific and commercial development at several institutions. These next-generation instruments have the potential of delivering the order- of-magnitude increases in DNA fragment analysis speed and throughput required for genomic research. However, to realize these potential benefits, advances in separation media are critically needed. The goal of this research is to develop a new, proprietary separation media technology with enabling features for narrow channel electrophoretic methods, such as capillary array electrophoresis. This new media technology will combine the best features of crosslinked gels and polymer solutions, i.e., high resolution and replaceability. The Phase I objectives are to: (1) Develop replaceable matrices for capillary electrophoresis of dsDNA with resolution equal to or greater than crosslinked polyacrylamide; and (2) Demonstrate the feasibility of separating sequencing fragments with single base resolution in replaceable matrices of the type proposed here. In Phase II, the sequencing matrix will be optimized to equal or surpass the resolution and read length of crosslinked polyacrylamide. PROPOSED COMMERCIAL APPLICATION: the replaceable gels developed in this research will have immediate commercial utility as an enabling component of narrow channel DNA electrophoresis instruments, such as capillary array electrophoresis. This project will thus contribute significantly to achieving the improvements in DNA fragment analysis speed and throughput required by the Human Genome Project.